Development of a cell-based nonradioactive glucose uptake assay system for SGLT1 and SGLT2

Kanwal, Abhinav; Singh, Shailendra Pratap; Grover, Paramjit; Banerjee, Sanjay K.

doi:10.1016/j.ab.2012.07.003

Cited by 44 publications

(29 citation statements)

References 17 publications

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“…Indeed, in some cases inhibition of glucose transport at high concentrations of chamomile or green tea reached ∼90%. When deoxy‐D‐glucose was used as the test substrate, substantial inhibition was observed, since this substrate is not metabolized further after entry into the cell, and is transported by SGLT1 and GLUT2, but not GLUT5 . In our models we confirmed that the drug acarbose does not inhibit any of the transport components tested, which supports the hypothesis that it could act synergistically with the chamomile or green tea on carbohydrate digestive processes in vivo (Figure ), and the work presented here opens the door to further in vivo studies.…”

Section: Discussionsupporting

confidence: 83%

Green and Chamomile Teas, but not Acarbose, Attenuate Glucose and Fructose Transport via Inhibition of GLUT2 and GLUT5

Villa-Rodríguez

Aydin

Gauer

et al. 2017

Molecular Nutrition Food Res

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Section: Discussionsupporting

confidence: 83%

Green and Chamomile Teas, but not Acarbose, Attenuate Glucose and Fructose Transport via Inhibition of GLUT2 and GLUT5

Villa-Rodríguez

Aydin

Gauer

et al. 2017

Molecular Nutrition Food Res

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“…To further investigate SGLT function in myocytes from diabetic and control rats, we measured glucose uptake using a fluorescent glucose analog, 2‐NBDG (Figure ) . 2‐NBDG is taken up into cells through both Glut transporters and SGLT . Glucose uptake was measured under control conditions, with SGLT blocked by 250 μmol/L phlorizin, and in the presence of 50 nmol/L insulin (Figure ).…”

Section: Resultsmentioning

confidence: 99%

Intracellular Na ⁺ Concentration ([Na ⁺ ] _i ) Is Elevated in Diabetic Hearts Due to Enhanced Na ⁺ –Glucose Cotransport

Lambert

Srodulski

Peng

et al. 2015

JAHA

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BackgroundIntracellular Na+ concentration ([Na+]i) regulates Ca2+ cycling, contractility, metabolism, and electrical stability of the heart. [Na+]i is elevated in heart failure, leading to arrhythmias and oxidative stress. We hypothesized that myocyte [Na+]i is also increased in type 2 diabetes (T2D) due to enhanced activity of the Na+–glucose cotransporter.Methods and ResultsTo test this hypothesis, we used myocardial tissue from humans with T2D and a rat model of late-onset T2D (HIP rat). Western blot analysis showed increased Na+–glucose cotransporter expression in failing hearts from T2D patients compared with nondiabetic persons (by 73±13%) and in HIP rat hearts versus wild-type (WT) littermates (by 61±8%). [Na+]i was elevated in HIP rat myocytes both at rest (14.7±0.9 versus 11.4±0.7 mmol/L in WT) and during electrical stimulation (17.3±0.8 versus 15.0±0.7 mmol/L); however, the Na+/K+-pump function was similar in HIP and WT cells, suggesting that higher [Na+]i is due to enhanced Na+ entry in diabetic hearts. Indeed, Na+ influx was significantly larger in myocytes from HIP versus WT rats (1.77±0.11 versus 1.29±0.06 mmol/L per minute). Na+–glucose cotransporter inhibition with phlorizin or glucose-free solution greatly reduced Na+ influx in HIP myocytes (to 1.20±0.16 mmol/L per minute), whereas it had no effect in WT cells. Phlorizin also significantly decreased glucose uptake in HIP myocytes (by 33±9%) but not in WT, indicating an increased reliance on the Na+–glucose cotransporter for glucose uptake in T2D hearts.ConclusionsMyocyte Na+–glucose cotransport is enhanced in T2D, which increases Na+ influx and causes Na+ overload. Higher [Na+]i may contribute to arrhythmogenesis and oxidative stress in diabetic hearts.

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“…Glucose uptake was measured in fully differentiated C2C12 cells using a non-radioactive fluorometric assay, as previously described (Leira et al, 2002; Zou et al, 2005; Kanwal et al, 2012). Briefly, following differentiation in 6 well plates, cells were serum starved in growth media for 2 h and cells were treated with either vehicle control, IL-15 100 ng/ml, insulin (100 nM), a STAT3 inhibitor (100 μM; S31-201; Sigma), or IL-15 + S31-20 with or without 50 μM 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG) (Cayman Chemical) in Krebs-Henseleit buffer containing 0.1% fatty acid free BSA (Sigma) and 2000 mg/L of glucose at 37°C with 5% CO 2 (Siddiquee et al, 2007; Kelly et al, 2010).…”

Section: Methodsmentioning

confidence: 99%

IL-15 Activates the Jak3/STAT3 Signaling Pathway to Mediate Glucose Uptake in Skeletal Muscle Cells

2016

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Myokines are specialized cytokines that are secreted from skeletal muscle (SKM) in response to metabolic stimuli, such as exercise. Interleukin-15 (IL-15) is a myokine with potential to reduce obesity and increase lean mass through induction of metabolic processes. It has been previously shown that IL-15 acts to increase glucose uptake in SKM cells. However, the downstream signals orchestrating the link between IL-15 signaling and glucose uptake have not been fully explored. Here we employed the mouse SKM C2C12 cell line to examine potential downstream targets of IL-15-induced alterations in glucose uptake. Following differentiation, C2C12 cells were treated overnight with 100 ng/ml of IL-15. Activation of factors associated with glucose metabolism (Akt and AMPK) and known downstream targets of IL-15 (Jak1, Jak3, STAT3, and STAT5) were assessed with IL-15 stimulation. IL-15 stimulated glucose uptake and GLUT4 translocation to the plasma membrane. IL-15 treatment had no effect on phospho-Akt, phospho-Akt substrates, phospho-AMPK, phospho-Jak1, or phospho-STAT5. However, with IL-15, phospho-Jak3 and phospho-STAT3 levels were increased along with increased interaction of Jak3 and STAT3. Additionally, IL-15 induced a translocation of phospho-STAT3 from the cytoplasm to the nucleus. We have evidence that a mediator of glucose uptake, HIF1α, expression was dependent on IL-15 induced STAT3 activation. Finally, upon inhibition of STAT3 the positive effects of IL-15 on glucose uptake and GLUT4 translocation were abolished. Taken together, we provide evidence for a novel signaling pathway for IL-15 acting through Jak3/STAT3 to regulate glucose metabolism.

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Development of a cell-based nonradioactive glucose uptake assay system for SGLT1 and SGLT2

Cited by 44 publications

References 17 publications

Green and Chamomile Teas, but not Acarbose, Attenuate Glucose and Fructose Transport via Inhibition of GLUT2 and GLUT5

Green and Chamomile Teas, but not Acarbose, Attenuate Glucose and Fructose Transport via Inhibition of GLUT2 and GLUT5

Intracellular Na ⁺ Concentration ([Na ⁺ ] _i ) Is Elevated in Diabetic Hearts Due to Enhanced Na ⁺ –Glucose Cotransport

IL-15 Activates the Jak3/STAT3 Signaling Pathway to Mediate Glucose Uptake in Skeletal Muscle Cells

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Development of a cell-based nonradioactive glucose uptake assay system for SGLT1 and SGLT2

Cited by 44 publications

References 17 publications

Green and Chamomile Teas, but not Acarbose, Attenuate Glucose and Fructose Transport via Inhibition of GLUT2 and GLUT5

Green and Chamomile Teas, but not Acarbose, Attenuate Glucose and Fructose Transport via Inhibition of GLUT2 and GLUT5

Intracellular Na + Concentration ([Na + ] i ) Is Elevated in Diabetic Hearts Due to Enhanced Na + –Glucose Cotransport

IL-15 Activates the Jak3/STAT3 Signaling Pathway to Mediate Glucose Uptake in Skeletal Muscle Cells

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Product

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Intracellular Na ⁺ Concentration ([Na ⁺ ] _i ) Is Elevated in Diabetic Hearts Due to Enhanced Na ⁺ –Glucose Cotransport